Synthesis and evaluation of monoclonal antibody against Plasmodium falciparum merozoite surface antigen 2

Objective:To assess the quality of expressed MSP-2 and also to confirm the immune response against different domains of these proteins.Methods:Mice were immunized with a schizont extract to stimulate the immune system to make antibodies against different antigens of the late stage parasite including production of antibodies against different domains of Plasmodium falciparum(P.falciparum)MSP-2.B lymphocytes of immunized mice were extracted from the spleen and the fusion was performed using NS-1 myeloma cells and the hybridoma cells were assayed by ELISA either with a schizont extract or different domains of MSP-2 and/or by IFAT with whole schizont preparation.Fusion of NS-1 and spleen cells was performed.The positive hybrids were cloned and ELISA was applied against different dilutions.The positive clones were transferred to a small tissue culture flask and after developing they were assayed against schizont extract and the different MSP-2 domains.The positive clones were expanded to large(75 cm^2)flask and cultured under the same conditions,checking them using both ELISA and IFAT and the positive cells were frozen as soon as possible.Results:A total number of 7 fusions including 26 plates(2496 wells)were performed,of which 1336 hybrids were produced and the overall efficiency(1336/24%×100)was about 53%.ELISA was performed to detect the positive hybrids against crude schizont extract by which the highest frequency to crude schizont extract was found for the supernatant of the hybrids produced in fusion number 3(66 out of 315 hybrids).The supernatant of both B5 and Ft hybridoma cells were more positive against domain 2 of the MSP-2 recombinant protein in Western blotting test.Western blotting results also showed that different domains of the MSP-2 recombinant protein and also the MSP-2 of the P.falciparum parasite were recognized by some of the positive clones and also immune sera.Conclusions:Bringing together all the results of this study it has been confirmed that some clones have recognized both schizont extract and different domains of the MSP-2 recombinant protein and therefore confirming the quality of the MSP-2 domains.

Etiology and resistance of asymptomatic bacteriuria isolates among school going children in Sanadaj-Iran

Objective:The objective of this study was to determine the etiology and antimicrobial susceptibility patterns of asymptomatic bacteriuria pathogens isolated among school going children in Sanandaj,Iran.Methods:A total of 1 457 urine samples of 5 to 10 years children from forty different schools of Sanandaj city were screened to see asymptomatic bacteriuria during November 2007 to June 2008.Results:Bacterial colony count of over (105) colony forming units CFU/ mL were found in 28(1.90%) of total cases,with 767(52.64%) girls and 690(47.35%) boys.The highest class-specific prevalence was found in the fourth standard(2.8%) and the lowest in the first standard(1.0%).The dominant bacterial isolates were E.coli 20(74.1%),followed by Klebsiella pneumoniae 04(14.8%) and Staphylococcus aureus 04(14.8%).Cefotaxime,Cefixime,Kanamycin, Co-trimoxazole,nalidicxic acid,nitrofurantoin and Amoxicillin,resistance rates were above 90.0%. Ceftriaxone expressed the highest susceptibility among E.coli isolates.Surprisingly,S.aureus showed 100. 0%resistance to oxacillin.Conclusion:In the present study in which Escherichia coli is the most frequently incriminated as the causative agents.The results show a very serious antibiotic resistance of E.coli isolated. Surveillance and monitoring studies will be essential in preventing of renal scarring or other abnormalities.